Potential of Lunasin Orally-Administered in Comparison to Intraperitoneal Injection to Inhibit Colon Cancer Metastasis in Vivo

Vermont P. Dia, Elvira Gonzalez de Mejia

Department of Food Science and Human Nutrition, University of Illinois at Urbana-Champaign, Urbana, USA.

DOI: 10.4236/jct.2013.46A2005                

Abstract

Lunasin is a bioactive peptide originally isolated from soybean and has demonstrated chemopreventive and anti-cancer properties against skin, colon and breast cancers. The objective of the study was to evaluate the capability of intraperitoneally and orally-administered soybean-derived peptide lunasin to inhibit KM12L4 human colon cancer cell metastasis in a mouse model. Intraperitoneal (i.p.) injection of lunasin (4 mg/kg bw/day) reduced the number of liver metastasis by 50% (P = 0.047) and the liver weight/body weight ratio by 23% (P = 0.039). Oral administration of lunasin reduced the number of liver metastasis by 56% (8 mg/kg bw/day, P = 0.293) and 94% (20 mg/kg bw/day, P = 0.247). Immunohistochemical staining of the liver-tissue section showed that lunasin at 20 mg/kg bw dose did not significantly reduce the expression of proliferating cell nuclear antigen, increased the expression of proapoptotic Bax by 2.7-fold but also increased the expression of the antiapoptotic Bcl-2 by 3.8-fold. Regarding epigenetic markers, H3K18 was not significantly affected by either oral dose while H4K8 was dose dependently increased by 2.3-fold (P = 0.001, 8 mg/kg bw) and 2.7-fold (P < 0.001, 20 mg/kg bw). On the other hand, i.p. injection of lunasin reduced both histone acetylation markers significantly. The difference on the effects can be attributed to the different routes of administration used leading to digestion of lunasin when given by oral gavage. In conclusion, lunasin reduced colon cancer metastasis in vivo; however, more studies are needed to determine the oral dose of lunasin and prevent colon cancer metastasis.

Keywords

Lunasin; Colon Cancer; Metastasis; Apoptosis; Histones

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	F. Bray, J. S. Ren, E. Masuyer and J. Ferlay, “Global Estimates of Cancer Prevalence for 27 Sites in the Adult Population in 2008,” International Journal of Cancer, Vol. 132, No. 5, 2013, pp. 1133-1145. doi:10.1002/ijc.27711
[2]	Y. D. Benoit, K. B. Laursen, M. S. Witherspoon, S. M. Lipkin and L. J. Gudas, “Inhibition of PRC2 Histone Methyltransferase Activity Increases TRAIL-Mediated Apoptosis Sensitivity in Human Colon Cancer Cells,” Journal of Cellular Physiology, Vol. 228, No. 4, 2013, pp. 764-772. doi:10.1002/jcp.24224
[3]	R. Wang, C. V. Lohr, K. Fischer, W. M. Dashwood, J. A. Greenwood, E. Ho, D. E. Williams, H. Ashtrokab, M. R. Dashwood and R. H. Dashwood, “Epigenetic Inactivation of Endothelin-1 And Endothelin-2 in Colon Cancer,” International Journal of Cancer, Vol. 132, No. 5, 2013, pp. 1004-1012. doi:10.1002/ijc.27762
[4]	L. G. Shack, A. Shah, P. C. Lambert and B. Rachet, “Cure by Age and Stage for Colorectal Cancer Patients in North West England, 1997-2004: A Population-Based Study,” Cancer Epidemiology, Vol. 36, No. 6, 2013, pp. 548-553.
[5]	R. R. Huxley, A. Ansary-Mogdahham, P. Clinton, S. Czernichow, C. L. Parr and M. Woodward, “The Impact of Dietary and Lifestyle Riskfactors on Risk of Colorectal Cancer: A Quantitative Overview of the Epidemiological Evidence,” International Journal of Cancer, Vol. 125, No. 1, 2009, pp. 171-180. doi:10.1002/ijc.24343
[6]	J. N. Poynter, S. B. Gruber, P. D. R. Higgins, R. Almog, J. D. Bonner, H. S. Rennert, M. Low, J. K. Greenson and G. Rennert, “Statins and the Risk of Colorectal Cancer,” New England Journal of Medicine, Vol. 352, No. 21, 2005, pp. 2184-2192.
[7]	M. R. Schweiger, M, Husoong, C, Rohr and H. Lehrach, “Genomics and Epigenomics of Colorectal Cancer,” Wiley Interdisciplinary Reviews: Systems Biology and Medicine, Vol. 5, No. 2, 2013, pp. 205-219.
[8]	L. Yan, E. L. Spitznagel and M. C. Bosland, “Soy Consumption and Colorectal Cancer Risk In Humans: A MetaAnalysis,” Cancer Epidemiology Biomarkers Prevention, Vol. 19, No. 1, 2010, pp. 148-158. doi:10.1158/1055-9965.EPI-09-0856
[9]	G. Yang, X. O. Shu, H. Li, W. H. Chow, H. Cai, X. Zhang, Y. T. Gao and W. Zheng, “Prospective Cohort Study of Soy Food Intake and Colorectal Cancer Risk in Women,” American Journal of Clinical Nutrition, Vol. 89, No. 2, 2009, pp. 577-583. doi:10.3945/ajcn.2008.26742
[10]	A. F. Galvez and B. O. de Lumen, “A Soybean cDNA Encoding a Chromatin-Binding Peptide Inhibits Mitosis of Mammalian Cells,” Nature Biotechnology, Vol. 17, No. 5, 1999, pp. 495-500.
[11]	H. J. Jeong, Y. Lam and B. O. de Lumen, “Barley Lunasin Suppresses Ras-Induced Colony Formation and Inhibits Core Histone Acetylation in Mammalian Cells,” Journal of Agricultural and Food Chemistry, Vol. 50, No. 21, 2002, pp. 5903-5908.
[12]	J. B. Jeong, H. J. Jeong, J. H. Park, S. H. Lee, J. R. Lee, H. K. Lee, G. Y. Chung and B. O. de Lumen, “CancerPreventive Peptide Lunasin from Solanum nigrum L. Inhibits Acetylation of Core Histones H3 and H4 and Phosphorylation or Retinoblastoma Protein (Rb),” Journal of Agricultural and Food Chemistry, Vol. 55, No. 26, 2007, pp. 10707-10713. doi:10.1021/jf072363p
[13]	H. J. Jeong, J. B. Jeong, D. S. Kim, J. O. Park, J. B. Lee, D. H. Kweon, G. Y. Chung, E. W. Seo and B. O. de Lumen, “The Cancer Preventive Peptide Lunasin from Wheat Inhibits Core Histone Acetylation,” Cancer Letters, Vol. 255, No. 1, 2007, pp. 42-48. doi:10.1016/j.canlet.2007.03.022
[14]	G. G. Meadows, “Diet, Nutrients, Phytochemicals, and Cancer Metastasis Suppressor Genes,” Cancer Metastasis Reviews, Vol. 31, No. 3-4, 2012, pp. 441-454. doi:10.1007/s10555-012-9369-5
[15]	B. Hernandez-Ledesma, C. C. Hsieh and B. O. de Lumen, “Relationships between Lunasin Sequence and Its Inhibitory Activity of Histones H3 and H4,” Molecular Nutrition and Food Research, Vol. 55, No. 7, 2011, pp. 989-998. doi:10.1002/mnfr.201000632
[16]	V. P. Dia and E. G. de Mejia, “Lunasin Promotes Apoptosis in Human Colon Cancer Cells by Mitochondrial Pathway Activation and Induction of Nuclear Clusterin Expression,” Cancer Letters, Vol. 295, No. 1, 2010, pp. 426-430.
[17]	V. P. Dia and E. G. de Mejia, “Lunasin Potentiates the Effect of Oxaliplatin Preventing Outgrowth of Colon Cancer Metastasis, Binds to Alpha 5 Beta 1 Integrin and Suppresses FAK/ERK/NF-Kappa B Signaling,” Cancer Letters, Vol. 313, No. 2, 2011, pp. 167-180. doi:10.1016/j.canlet.2011.09.002
[18]	K. Morikawa, S. M. Walker, M. Nakajima, S. Pathak and I. J. Fidler, “Influence of Organ Environment on the Growth, Selection and Metastasis of Human Colon Carcinoma Cell In Nude Mice,” Cancer Research, Vol. 49, No, 23, 1988, pp. 6863-6871.
[19]	A. Cavazos, E. M. Morales, V. P. Dia and E. G. de Mejia, “Analysis of Lunasin in Commercial and Pilot Plant Produced Soybean Products and an Improved Method of Lunasin Purification,” Journal of Food Science, Vol. 77, No. 5, 2012, pp. C539-C545. doi:10.1111/j.1750-3841.2012.02676.x
[20]	I. G. Zubeldia, A. M. Bleau, M. Redrado, D. Serrana, A. Agliano, C. Gil-Puig, F. Vinal-Vanaclocha, J. Lecanda and A. Calvo, “Epithelial to Mesenchymal Transition and Cancer Stem Cell Phenotypes Leading to Metastasis Are Abrogated by the Novel TGF-Beta-1 Peptides P17 and P144,” Experimental Cell Research, Vol. 319, No, 2013, pp. 12-22.
[21]	A. F. Galvez, N. Chen, J. Macasieb and B. O. de Lumen, “Chemopreventive Property of a Soybean Peptide (Lunasin) That Binds to Deacetylated Histones and Inhibits Acetylation,” Cancer Research, Vol. 61, No. 20, 2001, pp. 7473-7478.
[22]	C. C. Hsieh, B. Hernandez-Ledesma, H. J. Jeong, J. H. Park and B. O. de Lumen, “Complementary Roles in Cancer Prevention: Protease Inhibitor Makes the Cancer Preventive Peptide Lunasin Bioavailable,” Plos One, Vol. 5, No. 1, 2010, Article ID: e8890.
[23]	C. C. Hsieh, B. Hernandez-Ledesma and B. O. de Lumen, “Soybean Peptide Lunasin Suppresses in Vitro and in Vivo 7,12-Dimethylbenz-[a]Anthracene-Induced Tumorigenesis,” Journal of Food Science, Vol. 75, No. 9, 2010, pp. H311-H316. doi:10.1111/j.1750-3841.2010.01861.x
[24]	S. N. Narzhny, “Proliferating Cell Nuclear Antigen: A Proteomic View,” Cellular and Molecular Life Sciences, Vol. 65, No, 23, 2008, pp. 3789-3808. doi:10.1007/s00018-008-8305-x
[25]	S. Eltz, E. Comperat, O. Cussenot and M. Roupret, “Molecular and Histological Markers in Urothelial Carcinomas of the Upper Urinary Tract,” BJU International, Vol. 102, No. 5, 2008, pp. 532-535.
[26]	W. Zhong, J. Peng, H. He, D. Wu, Z. Han, X. Bi and Q. Dai, “Ki-67 and PCNA Expression in Prostate Cancer and Benign Prostatic Hyperplasia,” Clinical Investigations in Medicine, Vol. 31, No. 1, 2008, pp. E8-E15.
[27]	Z. Tan, M. Wortman, K. L. Dillehay, W. L. Seibel, C. R. Evelyn, S. J. Smith, L. H. Malkas, Y. Zheng, S. Lu and Z. Dong, “Small-Molecule Targeting of Proliferating Cell Nuclear Antigen Chromatin Association Inhibits Tumor Cell Growth,” Molecular Pharmacology. Vol. 81, No. 6, 2012, pp. 811-819. doi:10.1124/mol.112.077735
[28]	X. Z. Guo, X. D. Shao, M. P. Liu, J. H. Xu, L. N. Ren, J. J. Zhao, H. Y. Li and D. Wang, “Effect of bax, bcl-2 and bcl-xL on Regulating Apoptosis in Normal Liver and Hepatocellular Carcinoma,” World Journal of Gastroenterology, Vol. 8, No, 6, 2002, pp. 1059-1062.
[29]	B. Hrenandez-Ledesma, C. C. Hsieh and B. O. de Lumen, “Lunasin, a Novel Seed Peptide for Cancer Prevention,” Peptides, Vol. 30, No. 2, 2009, pp. 426-430. doi:10.1016/j.peptides.2008.11.002
[30]	V. P. Dia, S. Torres, B. O. de Lumen, J. W. Erdman and E. G. de Mejia, “Presence of Lunasin in Plasma of Young Men after Soy Protein Consumption,” Journal of Agricultural Food Chemistry, Vol. 57, No. 4, pp. 1260-1266.
[31]	H. M. Collins, M. K. Abdelghany, M. Messmer, B. Yue, S. E. Deeves, K. B. Kindle, K. MAntelingu, A. Aslam, G. S. Winkler, T. K. Kundu and D. M. Heery, “Differential Effects of Garcinol and Curcumin on Histone and p53 Modifications in Tumour Cells,” BMC Cancer, Vol. 13, 2013, Article ID: 37. doi:10.1186/1471-2407-13-37
[32]	Y. Wang and Y. Shang, “Epigenetic Control of Epithelial-to-Mesenchymal Transition and Cancer Metastasis,” Experimental Cell Research, Vol. 319, No. 2, 2013, pp. 160-169. doi:10.1016/j.yexcr.2012.07.019
[33]	J. S. Lee, E. Smith and A. Shilatifard, “The Language of Histone Crosstalk,” Cell, Vol. 142, No. 5, 2010, pp. 682-685. doi:10.1016/j.cell.2010.08.011
[34]	R. MArmorstein and S. Y. Roth, “Histone Acetyltransferase: Function, Structure, and Catalysis,” Current Opinion in Genetic Development, Vol. 11, No. 2, 2001, pp. 155-161.
[35]	Q. Li and H. Chen, “Epigenetic Modifications of Metastasis Suppressor Genes in Colon Cancer Metastasis,” Epigenetics, Vol. 6, No. 7, 2011, pp. 849-852. doi:10.4161/epi.6.7.16314
[36]	S. C. Truant, V. P. Gouyer, E. A. Leteurte, F. Zerimech, G. M. Huet and F. R. Pruvot, “E-Cadherin and BetaCatenin mRNA Levels throughout Colon Cancer Progression,” Journal of Surgical Research, Vol. 150, No. 2, 2008, pp. 212-218. doi:10.1016/j.jss.2007.12.800